A rapid nylon-fiber syringe system to deplete CD14+ cells for positive selection of human blood CD34+ cells. Use of immunomagnetic microspheres
A rapid nylon-fiber syringe system to deplete CD14+ cells for positive selection of human blood CD34+ cells. Use of immunomagnetic microspheres
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一种快速尼龙纤维注射器系统，用于消耗 CD14 细胞，以对人血液 CD34 细胞进行正选。
DOI：
10.1038/sj.bmt.1700659
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发表时间：
1997
影响因子：
4.8
通讯作者：
T. Koike
中科院分区：
文献类型：
--
作者：
M. Yamaguchi;K. Sawada;N. Sato;K. Koizumi;S. Sekiguchi;T. Koike
为了实现 CD34+ 细胞的快速有效纯化，我们设计了一种尼龙纤维注射器 (NF-S)，并操纵它来消除正常人血液单核细胞 (MN 细胞) 中的贴壁细胞。经NF-S处理的细胞进一步纯化为CD34+级分，使用CD34单克隆抗体、免疫磁性微球和木瓜凝乳蛋白酶处理来分离微球。当稳态人外周血MN细胞在24℃下经NF-S处理5分钟时，单核细胞（CD14+细胞）的频率从22.0±5.2%显着下降至2.5±0.4%，CD34+细胞的回收率为73%。随后的免疫磁性阳性选择获得了 91±8% 纯 CD34+ 细胞的制备物，产率为 86±23%。 CD34+细胞的总产量为44±11%，所有过程所需的时间为5小时。初始细胞群中 cd14+ 细胞的频率与最终制剂中 CD34+ 细胞的纯度之间存在紧密的负相关（P<0.0001）。为了获得超过 90% 纯 CD34+ 细胞的制剂，建议的 CD14+ 细胞频率低于 4.4%。当 NF-S 和免疫磁性微球相结合时，任何实验室都可以对稳态外周血中的 CD34+ 细胞进行高效的台式分离，并且不需要荧光激活细胞分选。
To achieve a rapid and an efficient purification of CD34+ cells, we devised a nylon-fiber syringe (NF-S) and we manipulated it to deplete adherent cells from normal human blood mononuclear cells (MN cells). The cells processed by NF-S were further purified as the CD34+ fraction, using CD34 monoclonal antibody, immunomagnetic microspheres and chymopapain treatment to detach the microspheres. When steady-state human peripheral blood MN cells were processed by NF-S at 24°C for 5 min, the frequency of monocytes (CD14+ cells) significantly decreased from 22.0 ± 5.2% to 2.5 ± 0.4%, with a 73% recovery of CD34+ cells. The subsequent immunomagnetic positive selection achieved preparations of 91 ± 8% pure CD34+ cells with a 86 ± 23% yield. The overall yield of CD34+ cells was 44 ± 11%, and the time required for all procedures was 5 h. There was a tight and an inverse correlation (P < 0.0001) between the frequency of cd14+ cells in the initial cell population and the purity of CD34+ cells in the final preparation. A recommended frequency of CD14+ cells for achieving preparations of over 90% pure CD34+ cells was less than 4.4%. When combining NF-S and immunomagnetic microspheres, efficient bench-top separation of CD34+ cells in steady-state peripheral blood can be done in any laboratory, and fluorescence- activated cell-sorting is not required.
乳腺癌或神经母细胞瘤患者输注 CD34+ 骨髓细胞后的植入。
影响因子：
20.3
作者：
Berenson,RJ;Bensinger,WI;Hill,RS;Andrews,RG;Garcia-Lopez,J;Kalamasz,DF;Still,BJ;Spitzer,G;Buckner,CD;Bernstein,ID
通讯作者：
Bernstein,ID